Journal: Nucleic acids research

Article Title: Cellular APOBEC3A deaminase drives mutations in the SARS-CoV-2 genome.

doi: 10.1093/nar/gkac1238

Figure Lengend Snippet: Figure 3. A3A mRNA expression in human airway and lung cells. (A) The A3A mRNA level in each cell line was quantified by RT–ddPCR 18 h after treatment without (control) or with IFN-ß and/or TNF- under normoxic conditions (N). A549, Calu-3, SAE, AT2 and LBE cells and HNEpCs were also cultured under hypoxic conditions (H). The mean mRNA copy num- bers of A3A relative to those of the housekeeping gene RPP40 are shown (n = 3). (B) The CD11b and CD68 mRNA levels in each untreated cell line were quantified by RT–ddPCR. Human MDMs were used as positive controls. The mean mRNA copy numbers of CD11b and CD68 relative to those of RPP40 are shown (n = 3). The error bars indicate the + SD values.

Article Snippet: Human nasal epithelial primary cells (HNEpCs) and human type II alveolar epithelial (AT2) cells were purchased from PromoCell and Accegen Biotechnology, respectively.

Techniques: Expressing, Control, Cell Culture

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Burkholderia mallei and Burkholderia pseudomallei stimulate differential inflammatory responses from human alveolar type II cells (ATII) and macrophages

doi: 10.3389/fcimb.2012.00165

Figure Lengend Snippet: B. mallei ( BM ) and B. pseudomallei ( BP ) interaction with primary human ATII cells (ATII cells). ATII cells were grown on 3.0-um transwell filters to confluence prior to apical exposure of BM and BP at an MOI of approximately 100:1 for 4 h. Minimal contact of BM ( A and B ) with ATII cells was visualized via SEM. However, BP (C–E) adhesion to the apical surface of ATII cells was observed and the pathogen was entangled by the ATII microvilli [ D (white boxed area) and E ]. Further, quantitative analysis of attachment showed that BP significantly adhered better to ATII cells compared to BM ( p < 0.05; F ). Data is representative of triplicate samples of three experiments and is represented as mean ± SEM. * p < 0.05.

Article Snippet: Primary human type II alveolar epithelial (ATII) cells (ScienCell, San Diego, CA) were seeded on BioCoat Growth Factor Reduced Matrigel®-coated transwell filters (BD; Franklin Lakes, NJ) and maintained using Bronchial Epithelial Medium enhanced with the Bullet kit supplements provided by the manufacturer (BEGM; Lonza, Basel, Switzerland) supplemented with 10% charcoal-stripped FBS (Hyclone; Logan, UT) and 10 ng/mL of keratinocyte growth factor (KGF; PeproTech, Rockhill, UT).

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Burkholderia mallei and Burkholderia pseudomallei stimulate differential inflammatory responses from human alveolar type II cells (ATII) and macrophages

doi: 10.3389/fcimb.2012.00165

Figure Lengend Snippet: BM and BP -induced time-dependent secretion of cytokines from ATII cells and MDM. ATII cells and MDM were infected with BM or BP (MOI 100:1) for 6 or 20 h. Supernatant was collected and analyzed for cytokine secretion. Un-stimulated cells served as baseline controls. BP stimulated higher levels of IL-6 (A) and TNF-α (B) from MDM as well as IL-6 (D) and IL-8 (E) from ATII cells at 6 and 20 h post-infection compared to BM . However, BM -induced significantly higher IL-10 secretion from MDM compared to BP at 6 h post-infection (C) . Additionally, IL-10 was significantly secreted from BM-ATII cells compared to BP at both 6 and 20 h post-infection (F) . Data is representative of two experiments containing duplicate conditions/experiment and is represented as mean ± SEM. * denotes significance of p < 0.05.

Article Snippet: Primary human type II alveolar epithelial (ATII) cells (ScienCell, San Diego, CA) were seeded on BioCoat Growth Factor Reduced Matrigel®-coated transwell filters (BD; Franklin Lakes, NJ) and maintained using Bronchial Epithelial Medium enhanced with the Bullet kit supplements provided by the manufacturer (BEGM; Lonza, Basel, Switzerland) supplemented with 10% charcoal-stripped FBS (Hyclone; Logan, UT) and 10 ng/mL of keratinocyte growth factor (KGF; PeproTech, Rockhill, UT).

Techniques: Infection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Burkholderia mallei and Burkholderia pseudomallei stimulate differential inflammatory responses from human alveolar type II cells (ATII) and macrophages

doi: 10.3389/fcimb.2012.00165

Figure Lengend Snippet: Phagocytosis and intracellular replication of BM and BP . MDM and ATII cells were infected with BM or BP (MOI of 100:1). As determined by CFUs, phagocytosis (accessed at 1 h) and intracellular replication (accessed at 3 h post-infection) of BP by MDM was significantly higher compared to BM (A) . Alternatively, neither BM nor BP invaded or replicated to high numbers in ATII cells (B) . Data is representative of two experiments containing duplicate conditions/experiment and is represented as mean ± SEM. * denotes significance of p < 0.05 and ** p < 0.01.

Article Snippet: Primary human type II alveolar epithelial (ATII) cells (ScienCell, San Diego, CA) were seeded on BioCoat Growth Factor Reduced Matrigel®-coated transwell filters (BD; Franklin Lakes, NJ) and maintained using Bronchial Epithelial Medium enhanced with the Bullet kit supplements provided by the manufacturer (BEGM; Lonza, Basel, Switzerland) supplemented with 10% charcoal-stripped FBS (Hyclone; Logan, UT) and 10 ng/mL of keratinocyte growth factor (KGF; PeproTech, Rockhill, UT).

Techniques: Infection

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Concept of the 3D lung-on-a-chip model based on biomimetically microcurved culture membranes. (A) We approached a structure similar to a cut and flipped open alveolar sac as a cell-populated membrane with the microcurved shape, size, and also arrangement of its bioartificial alveoli mimicking the ones of the adult organ. Integrated in microfluidic chips/OoC devices where the microcurved membranes separate a top from a bottom compartment, they can be seeded by infusion with lung epithelial and microvascular endothelial cells on the top and bottom side of the membrane. The spatial cell distribution is then similar to the alveolar–capillary barrier. (B) Potential future applications of the model include 3D ALI culture (following submerged culture), modeling of disease and repair/regeneration, and toxicity and pharmaceutical efficacy testing (temporarily under submerged conditions or exposed to vapors or aerosols).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Membrane

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Geometrical characterization of the (bottom) housing half of the chip from PDMS, and perfusion and leak test of the assembled OoC device. (A) Each housing half of the microfluidic chip contained one of the two central circular culture chambers with a diameter of 8 mm for receiving the hexagonal microwell array. This chamber was on either side connected to an inlet and an outlet channel with in each case a width of 500 μm and a length of 4 mm. At their lateral/peripheral ends across the culture chamber, the two channels in turn were connected to in each case one smaller chamber with a diameter of 1 mm located in two opposite corners of the chip. The height/depth of the microfluidic compartments was around 400 μm. (B) Cross-section of an assembled 3D lung-on-chip device (stitched image; housing halves that during cell culture host epithelial and endothelial cells are colored blue and pink/purple, respectively; scale bar represents 500 μm). (C) Assembled lung-on-chip device with its top and bottom chip compartment perfused through press-fitted tubing with water colored with green and blue (food) dye, respectively (scale bar represents 8 mm).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Cell Culture

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Epithelialization of the microcurved membrane in the chip. HAECs cultured submerged under flow for 7 days and stained for cell nuclei and (A) F-actin, (B) tight junctions, (C) vimentin, and (D) CK8 (fluorescent microscopy images; nuclei not shown in the right halves of the images for better visibility of the individual stains; scale bars represent 100 μm).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Membrane, Cell Culture, Staining, Microscopy

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: ALI culture on the microcurved membrane in the chip. HAECs cultured at the ALI under perfusion for 14 days and stained for cell nuclei and (A) F-actin, (B) CK8, (C) aquaporin 5, and (D) pSPC (fluorescent microscopy images; scale bars represent 100 μm).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Membrane, Cell Culture, Staining, Microscopy

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Thickness of the formed, curved alveolar epithelial layer. The thickness of the HAEC lining was measured (A) in two perpendicular cross-sections and there in each case in five different locations: at the horizontal center of the bottom of the microwell, at the left and right sidewall of the microwell directly under its convex rim, and at the left and right sidewall roughly halfway between, in each case perpendicular to the microwell wall. (B) Representative vertical and horizontal cross-sectional images of the epithelial layer (image planes “ x ” and “ y ”, and “ z 1 ” to “ z 3 ”, respectively; scale bars represent 50 μm). (C) Graph of the HAEC layer thickness as a function of the measurement location as stated in (A) ( n = 3).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques:

Journal: ACS Biomaterials Science & Engineering

Article Title: 3D Lung-on-Chip Model Based on Biomimetically Microcurved Culture Membranes

doi: 10.1021/acsbiomaterials.1c01463

Figure Lengend Snippet: Lung epithelial and endothelial coculture on the microcurved membrane in the chip. (A) Top views of sections of the microcurved membrane with Calu-3 cells cultured for 11 days and stained for cell nuclei and tight junctions (fluorescent microscopy image; scale bars represent 100 μm). (B) Bottom views of sections of the same microcurved membrane with HLMVECs cultured for 11 days and stained for nuclei and CD31 (fluorescent microscopy image; scale bars represent 100 μm). (C) Cross-section of the microcurved membrane from (A) and (B) (scale bar represents 100 μm). (D) Graph of the count of Calu-3 cells ( n = 4) and HLMVECs per square microwell unit ( n = 3) (**** indicates a p -value smaller than 0.0001).

Article Snippet: For the ALI culture, commercially available primary human alveolar epithelial cells (HAECs; PELOBiotech/Cell Biologics) were used.

Techniques: Membrane, Cell Culture, Staining, Microscopy